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The FACSCalibur flow cytometer is an autonomous benchtop flow cytometer designed for routine cell analysis, assay development, verification and identification of cellular populations. It is equipped with a blue (488 nm) air-cooled argon laser and a red (635 nm) diode laser. For each particle (cell), five optical parameters can be recorded from the 488 nm laser beam excitation: two light scatter signals, namely forward and right angle, and three fluorescences corresponding to emissions in green (530/30 nm BP), orange (585/42 nm BP) and red (670 nm LP) wavelength ranges. A far red fluorescence (661/16 nm BP) induced by the red diode can also be recorded. Data are analysed using BD Biosciences CellQuest software. Optional features include a cell sorting option, allowing users to identify and isolate a population of interest and a HTS option (High-throughput (HT) or Standard (STD) mode), where sample volumes range from 2-10 microlitres in HT mode and 2-200 microlitres in STD mode. An optional BD FACS Loader tube-lifter can be used to verify tube position and rack identification. The instrument has a capture rate of 300 cells per second, supports 40 (12 x 75 mm) tubes per rack, and has an operating temperature ranging from 16-29 degC.